The objective of this research proposal is to study the interaction of non-histone chromosomal proteins to DNA in chromatin. This involves the identification of unique binding sites for non-histones in DNA. This may be done by first fractionating DNA relative to its base sequence homology, then identify and later isolate unique non-histone proteins. The binding of non-histones to DNA may further be characterized by their specific effects on in vitro transcription of reconstituted chromatin. The in vitro experiments involve reconstitution of chromatin from one cell type to specific non-histones of a second cell type from the same species. This chromatin is transcribed and RNA base sequences examined by hybridization to DNA of unique sequences. Other experiments are also proposed to examine possible specific competitive binding of non-histones to DNA as they are obtained from two distinct and homologous chromatin. In this group of experimentation we will, therefore, be looking for specificity not only in the proteins but also in the DNA: (1) specific protein binding to DNA of different Cot values; and (2) Cot curve for DNA binding to specific fractions of NHCP. The methods entail the propagation of cleft palates in mice with a glucocorticoid and chromatin will then be isolated first from normal mouse brain and liver, and then from cleft palates of affected embryos. Physical methods, such as melting profiles, CD spectra and ethidium bromide binding to chromatin, will also be used to study effects of protein binding to DNA on the structure in DNA.